Part:BBa_K3505036

pFliC:RBS-LacI-Terminator- pAndersonJ23115:LacO:RBS-ECFP-terminator

Fig.1: Activation of eCFP in absence of SCFAs,Based on NOT GATE

Fig.2: The NOT GATE

Usage and Biology

2 Trancription Units

• LacI BBa_K3505003regulated by inducible promoter FliC BBa_K2924016which is activated from SFCAs and more specifically from Butyrate
• eCFP BBa_K3505019under control of lac regulated AndersonBBa_K3505013.

This way we take signal in the Absence of SCFAs

Design Notes

The coding sequence was domesticated . We removed BsmBI ,BsaI , BtgZI, BpiI sites in order to be compatible with GoldenBraid and MoClo. The sequence is present in BBa_K3505010 and has overhangs compatible for Golden Braid cloning.

Verification of cloning

Fig.3:U2 C2 pFliC:RBS-LacI-Terminator-pAndersonJ23115:LacO:RBS-ECFP-terminator EcoRV, HindIII. Expected bands 3734, 1699.

Experimental Use and Experience

Fig.3:J23115-eCFP-Terminator

Fig.4:J23115-eCFP-Terminator

Fig.5:J23115-eCFP-Terminator

Fig.6:J23115-eCFP-Terminator

Sequnce and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal NheI site found at 1321
  Illegal NheI site found at 1344
  
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]